Simultaneous quantification of estrogens and glucocorticoids in human adipose tissue by liquid-chromatography-tandem mass spectrometry

Sofia Laforest, Mélissa Pelletier, Nina Denver, Brigitte Poirier, Sébastien Nguyen, Brian R. Walker, Francine Durocher, Natalie Z.M. Homer, Caroline Diorio, André Tchernof, Ruth Andrew

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Abstract

The presence of estrogens, androgens and glucocorticoids as well as their receptors and steroid converting enzymes in adipose tissue has been established. Their contribution to diseases such as obesity, diabetes and hormone-dependent cancers is an active area of research. Our objective was to develop a LC–MS/MS method to quantify bioactive estrogens and glucocorticoids simultaneously in human adipose tissue. Estrogens and glucocorticoids were extracted from adipose tissue samples using solid-phase extraction. Estrogens were derivatized using 1-(2,4-dinitro-5-fluorophenyl)-4-methylpiperazine (PPZ) and methyl iodide to generate a permanently charged molecule (MPPZ). Steroids were separated and quantified by LC–MS/MS. The limit of quantitation for the steroids was between 15 and 100 pg per sample. Accuracy and precision were acceptable (<20%). Using this method, estradiol, estrone, cortisone and cortisol were quantified in adipose tissue from women with and without breast cancer. This novel assay of estrogens and glucocorticoids by LC–MS/MS coupled with derivatization allowed simultaneous quantification of a panel of steroids in human adipose tissue across the endogenous range of concentrations encountered in health and disease.

Original languageEnglish
Article number105476
Number of pages9
JournalJournal of Steroid Biochemistry and Molecular Biology
Volume195
Early online date24 Sep 2019
DOIs
Publication statusPublished - 31 Dec 2019

Keywords

  • estradiol
  • estrone
  • cortisol
  • cortisone
  • derivatization
  • adipose

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    Laforest, S., Pelletier, M., Denver, N., Poirier, B., Nguyen, S., Walker, B. R., Durocher, F., Homer, N. Z. M., Diorio, C., Tchernof, A., & Andrew, R. (2019). Simultaneous quantification of estrogens and glucocorticoids in human adipose tissue by liquid-chromatography-tandem mass spectrometry. Journal of Steroid Biochemistry and Molecular Biology , 195, [105476]. https://doi.org/10.1016/j.jsbmb.2019.105476